Multiplexed Nucleic Acid Programmable Protein Arrays

Yu, Xiaobo; Song, Lusheng; Petritis, Brianne; Bian, Xiaofang; Wang, Haoyu; Viloria, Jennifer; Park, Jin; Bui, Hoang; Li, Han; Wang, Jie; Liu, Lei; Yang, Liuhui; Duan, Hu; McMurray, David N.; Achkar, Jacqueline M.; Magee, Mitch; Qiu, Ji; LaBaer, Joshua

We compared the protein display between the two array types using groups of five proteins with either similar (Figure 2A and Figure 2B ) or varied molecular weights (Figure 2A and Figure 2C) , and then calculated the signal ratio of M-NAPPA to NAPPA. In both cases, all of the antibodies readily detected their corresponding antigens. For the spots with similarly-sized proteins (36 kDa to 85 kDa), the signal ratio of M-NAPPA to NAPPA was 0.78±0.44. For the spots containing five proteins covering a wide range of molecular weight, from 29 kDa to 106 kDa (Figure 2C) , the binding signal ratio of M-NAPPA to NAPPA was 1.03±0.75 ( Figure S3) . Thus, multiplexing proteins of similar size did not confer any advantage over random multiplexing. To further demonstrate that there were no biases in the expression of different proteins produced from mixed plasmids, five-plasmid mixtures containing various combinations of seven different genes (Abl1, IA-2, GAD2, Jun, RhoU, BCL2L2 and MT3033) were co-expressed in IVTT solution and analyzed via western blot. Despite a wide range of protein sizes, all proteins were expressed at similar amounts in their relevant combinations ( Figure S4) .